Award details

Structure and function of a novel gammaherpesvirus determinant of pathogenesis

Reference	BB/C507337/1
Principal Investigator / Supervisor	Professor Bernadette Dutia
Co-Investigators / Co-Supervisors	Professor Alastair Aitken, Professor Anthony Nash, Professor Malcolm Walkinshaw
Institution	University of Edinburgh
Department	Veterinary Biomedical Sciences
Funding type	Research
Value (£)	207,714
Status	Completed
Type	Research Grant
Start date	01/11/2004
End date	31/10/2007
Duration	36 months

Abstract

Members of the gammaherpesvirus family encode immunomodulatory molecules that enable viruses to establish and maintain latent infections in the presence of the host immune system. Murine gammaherpesvirus 68 (MHV-68) encodes 4 novel proteins, M1-M4. M3 ORF encodes a chemokine binding protein which alters virus pathogenicity and recently we have shown that the product of the M4 ORF plays a role in the modulating innate immune response. In this grant we propose to investigate the precise function of the M4 protein. We will express the M4 protein in the baculovirus system and purify mg quantities of the protein. The protein will be characterised by gel electrophoresis, circular dichroism and mass spectrometry. We will raise antisera in rabbits to the recombinant protein and carry out expression studies on M4. We will use the recombinant protein to identify cells and/or molecules with which M4 interacts. Murine splenocytes binding M4 will be purified with anti-M4 antibody by MACS and identified by FACS analysis. Molecules that bine M4 will be purified with anti-M4 antibody by MACS and identified by FACS analysis. Molecules that bind M4 will be identified by affinity purification and fractionated on 1D and 2D gels. Proteins will be identified by tryptic digestion followed by MALDI-TOF. Functional pathways will be characterised by 2D gel analysis. The M4 protein will be crystallized and using the hanging drop vapour diffusion method or batch method and the 3D structure determined.

Summary

unavailable

Committee	Closed Committee - Animal Sciences (AS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Proteomics and Cell Function (PCF) [2003-2004]
Funding Scheme	X – not Funded via a specific Funding Scheme

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